2004
CKI1, a putative sensor histidine kinase is essential for proper female gametophyte formation in Arabidopsis thaliana.
HEJÁTKO, Jan, Markéta PERNISOVÁ, Tinka ENEVA, Klaus PALME, Břetislav BRZOBOHATÝ et. al.Základní údaje
Originální název
CKI1, a putative sensor histidine kinase is essential for proper female gametophyte formation in Arabidopsis thaliana.
Název česky
CKI1, predpokladana receptorova histidinkinaza je nezbytna pro normalni vyvoj samiciho gametofytu u Arabidopsis thaliana.
Autoři
HEJÁTKO, Jan (203 Česká republika), Markéta PERNISOVÁ (203 Česká republika), Tinka ENEVA (100 Bulharsko), Klaus PALME (276 Německo) a Břetislav BRZOBOHATÝ (203 Česká republika, garant)
Vydání
Hradec Králové, Boook of Abstracts, Cytokinematics 2004, Microscopy of Live Cells in the Post Genomics Era. The 8th Symposium on Light Microscopy & Live Cells in Hradec Kralove, s. 22-22, 2004
Nakladatel
RNDr. F. Skopec, CSc.-NUCLEUS HK
Další údaje
Jazyk
angličtina
Typ výsledku
Stať ve sborníku
Obor
Genetika a molekulární biologie
Stát vydavatele
Německo
Utajení
není předmětem státního či obchodního tajemství
Odkazy
Kód RIV
RIV/00216224:14310/04:00010681
Organizační jednotka
Přírodovědecká fakulta
ISBN
80-86225-60-7
Klíčová slova anglicky
female gametophyte development; two-component signaling; sensor histidine kinase; early seed development; genomic imprinting
Štítky
Změněno: 28. 4. 2006 17:31, doc. RNDr. Jan Hejátko, Ph.D.
V originále
Embryo sac formation is a fundamental step in plant sexual reproduction. However, the key players driving female gametophyte development remain elusive. Employing reverse genetics, we have identified CKI1, a putative sensor histidine kinase, as a key regulatory component during female gametophyte formation in Arabidopsis thaliana. Using PCR-based screen we have identified a line carrying insertion of the autonomous transposable element En-1 in the coding region of CKI1 gene. Distorted genotype ratios and distribution of the insertional mutant allele cki1-i in the progeny of the identified mutant plant suggested lowered viability and/or function of the cki1-i gametes. In accordance to that, the results of reciprocal back-crosses have shown that cki1-i cannot be transmitted through the female germ line. Confocal laser scanning microscopy (CLSM) analysis of developing embryo sacs from cki1-i/CKI1 pistils did not reveal any deviations from the wild type phenotype in the first four developmental stages of megagametogenesis, FG1 through FG4. First anatomically distinguishable defects (distortion of the central vacuole, changes in the nuclei positions and partial degradation of the embryo sac) were identified in late FG5. The results of CLSM analysis 24 hours after flower emasculation (HAE) have confirmed that half of the embryo sacs from cki1-i/CKI1 pistils are unable to develop beyond the FG5. CLSM analysis 48 HAE revealed dynamic nature of the progressive degradation of mutant embryo sacs, very probably as a result of the central vacuole disintegration. The transcriptional activity of CKI1 during megagametogenesis was identified using in situ CKI1 mRNA analysis on paraffin sections of the developing embryo sacs. We have found weak but distinct signal throughout the embryo sac development, suggesting very early transcriptional activation of CKI1 during megagametogenesis. The results were confirmed by the histochemical analysis of stable transformants carrying fusion of the CKI1 promoter region with marker gene uidA. The GUS activity in developing embryo sacs was detectable as early as in the FG1 and remained detectable throughout megagametogenesis, even after the embryo sacs reached complete maturity at FG7. CKI1 expression was found not only just before but also early after fertilization in developing seed endosperm, which might suggest the role of CKI1 also in a newly formed sporophytic tissue. Surprisingly, transcriptional activation of the paternally transmitted uidA gene under control of CKI1 promoter very early after fertilization suggests potential transcriptional activity of at least part of the male genome during early seed development in Arabidopsis.
Česky
Tvorba embryonalniho vaku je nezbytnou soucasti pohlavni reprodukce rostlin. Presto zustavaji zakladni faktory ridici tvorbu samiciho gametofytu nezname. Za pouziti reverzni genetiky jsme identifikovali CKI1, predpokladanou receptorovou histidinkinazu jako klicovou regulacni komponentu behem tvorby samiciho gametofytu u Arabidopsis thaliana. Pomoci PCR vyhledavani jsme identifikovali lini nesouci inzerci kukuricneho autonomniho elementu En-1 v kodujici oblasti genu CKI1. Zmena v segregacnich pomerech a distribuce inzercni mutantni alely cki1-i v potomstvu identifikovane mutantni rostliny nazancovaly snizenou zivotnost, resp. funkcnost gamet nesoucich mutantni alelu cki1-i. Ve shode s tim, vysledky zpetnych reciprokych krizeni potvrdily, ze alela cki1-i nemuze byt do dalsiho potomstva prenesena prostrednictvim samici zarodecne linie. Analyza embryonalnich vaku u rostlin cki1-i/CKI1 pomoci laserove konfokalni mikroskopie (CLSM), ukazala normalni vyvoj vsech embryonalnich vaku do stadia FG4. Prvni anatomicky odlisitelne odchylky od normalniho vyvoje byly identifikovany teprve v pozdnim stadiu FG5. CLSM analyza 24 hodin po kvetni emaskulaci (HAE)potvrdila, ze polovina z embryonalnich vaku v rostlinach cki1-i/CKI1 je neschopna dokoncit stadium FG5. Vysledky teto analyzy 48 HAE prakazaly dynamickou povahu progresivni degradace muatnich embryonalnich vaku, pravdepodobne jako vysledek rozpadu centralni vakuoly. Aktvita transkripce genu CKI1 behem samici gametogeneze byla identifikovana pomoci in situ lokalizace CKI1 mRNA na parafinovych rezech. Pritomnost slabeho signalu byla zjistena behem celeho prunehu megagametogeneze, a to od velmi rannych stadii. Transkripcni aktivita genu CKI1 od sameho pocatku megagametogeneze byla potvrzena pomoci histochemicke analyzy stabilnich transformantnich linii nesoucich markrovy gen uidA pod kontrolou promotorove oblasti genu CKI1. GUS aktivita byla identifikovatelna od stadia FG1 a dale behem cele megagametogeneze. Exprese CKI1 byly identifikovana nejen tesne pred ale i po oplozeni v endospermu vyvijejiciho se semene, coz by mohlo znamenat potencialni roli CKI1 v nove vytvorenem sporofytu. Prekvapive byla transkripcni aktivita paternalni alely CKI1 identifikovana velmi brzy po oplozeni, coz nanznacuje aktivaci alespon casti parentalniho genomu behem ranych stadii vyvoje semene u Arabidopsis.
Návaznosti
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